Metabolism of the dimethyl ester of [2,3-13C]succinic acid in rat hepatocytes |
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Authors: | Malaise WJ Ladrière L Jijakli H Laatikainen R Niemitz M Verbruggen I Biesernans M Willem R |
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Institution: | (1) Laboratory of Experimental Medicine, Brussels Free University (ULB), Brussels, Belgium;(2) Kuopio NMR Research Group, Department of Chemistry, University of Kuopio, Kuopio, Finland;(3) High Resolution NMR Centre, Free University of Brussels (VUB), Brussels, Belgium |
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Abstract: | Hepatocytes prepared from overnight fasted rats were incubated for 120 min in the presence of the dimethyl ester of 2,3-13C]succinic acid (10 mM). The identification and quantification of 13C-enriched metabolites in the incubation medium were performed by a novel computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints. The generation of 13C-labelled metabolites, including succinate, fumarate, malate, lactate, alanine, aspartate and glucose accounted for about half of the initial amount of the ester present in the incubation medium. A fair correlation was observed between the experimental abundance of each 13C-labelled glucose isotopomer and the corresponding values derived from a model for the metabolism of 2,3-13C]succinate. Newly formed glucose was more efficiently labelled in the carbon C5 than C2, as well as the carbon C6 than C1, supporting the concept that D-glyceraldehyde-3-phosphate may undergo enzyme-to-enzyme channelling between glyceraldehyde-3-phosphate dehydrogenase and phosphofructoaldolase. |
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Keywords: | rat hepatocytes succinic acid dimethyl ester gluconeogenesis |
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